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Addgene
Programmable protein delivery with a bacterial contractile injection system.
Kreitz J, Friedrich MJ, Guru A, Lash B, Saito M, Macrae RK, Zhang F
Nature. 2023 Apr;616(7956):357-364. doi: 10.1038/s41586-023-05870-7. Epub 2023 Mar 29. PubMed Article

Plasmids from Article

ID Plasmid Purpose
198271pAWP78-PVCpnf1-16Unmodified pPVC; contains WT PVC structural/accessory genes
198272pBR322-PVCpnf17-22Unmodified pPayload; contains WT PVC payloads/regulatory genes
198273pBR322-PVCpnf18-21Contains only regulatory genes; for generating empty PVCs
198274pBR322-Pdp1_NTD-CreFor loading Cre into PVCs; also contains PVC regulatory genes
198275pBR322-Pdp1_NTD-Cas9For loading SpCas9 into PVCs; also contains PVC regulatory genes
198276pBR322-Pdp1_NTD-ZFD_LFor loading L half of zinc finger deaminase targeting human TRAC locus (ZFD_TRAC-NC-Left-G1397-N_DddA); also contains PVC regulatory genes
198277pBR322-Pdp1_NTD-ZFD_RFor loading R half of zinc finger deaminase targeting human TRAC locus (ZFD_TRAC-NC-Right-G1397-C_DddA); also contains PVC regulatory genes
198278pIEx4-Dio-GFPCre reporter for insect cells; double-floxxed-EGFP
198279pcDNA3-Dio-GFPCre reporter for mammalian cells; double-floxxed-EGFP
198280pIEx4-CreExpresses Cre in insect cells
198281pcDNA3-CreExpresses Cre in mammalian cells
198282py100-sgRNA_hVEGFAExpression of SpCas9 guide RNA specific for human VEGFA locus
198283pAWP78-PVCpnfΔpvc13Produces PVCs lacking tail fibers
198284pAWP78-PVCpnfΔpvc15Produces PVCs lacking payloads by elimination of the payload loader (pvc15)
198285pAWP78-PVCpnf_FLAG-pvc2Contains an external epitope tag on the sheath protein; for immunofluorescence-based binding assays
198286pAWP78-PVCpnf_pvc13-truncatedContains a tail fiber with a truncated binding domain (lacking aa403-476 in Pvc13); produces nontargeting PVCs
198287pAWP78-PVCpnf_pvc13-E01DARPinProduces PVCs retargeted with anti-EGFR DARPin E01
198288pAWP78-PVCpnf_pvc13-A4DARPinProduces PVCs retargeted with anti-lysozyme DARPin A4
198289pAWP78-PVCpnf_pvc13-Ad5KnobProduces PVCs retargeted with knob domain from hAd5 virus
198290pAWP78-PVCpnf_pvc13-Ad5DL491-492Produces PVCs retargeted with non-binding mutant of Ad5 knob
198291pAWP78-PVCpnf_pvc13-Ad5RGDPK7Produces PVCs retargeted with expanded tropism hAd5 knob (RGD and PK7 insertions)
198292pAWP78-PVCpnf_pvc13-antiMouseMHCIINbProduces PVCs retargeted with VHH7 anti-mouse MHC class II nanobody
198293pAWP78-PVCpnf_pvc13-antiHistoneNbProduces PVCs retargeted with A9 anti-histone nanobody
198294pAWP78-PVCpnf_pvc13-CD4DARPinProduces PVCs retargeted with anti-CD4 DARPin D27.2
198295pAWP78-PVCpnf_pvc13-CD11bDARPinProduces PVCs retargeted with anti-CD11b DARPin F7
198296pAWP78-PVCpnf_pvc13-HAProduces PVCs retargeted with HA tag
198297pAWP78-PVCpnf_pvc13-FLAGProduces PVCs retargeted with FLAG tag
198298pAWP78-PVCpnf_pvc13-EEProduces PVCs retargeted with EE tag
198299pAWP78-PVCpnf_pvc13-MoonTagProduces PVCs retargeted with MoonTag
198300pAWP78-PVCpnf_pvc13-SunTagProduces PVCs retargeted with SunTag
198301pAWP78-PVCpnf_pvc13-ALFAProduces PVCs retargeted with ALFA tag
198302pHCMV-SS-antiHAscFv-TMDisplays anti-HA scFv from the surface of mammalian cells
198303pHCMV-SS-antiFLAGscFv-TMDisplays anti-FLAG scFv from the surface of mammalian cells
198304pHCMV-SS-antiEEscFv-TMDisplays anti-EE scFv from the surface of mammalian cells
198305pHCMV-SS-antiMoonTagNb-TMDisplays anti-MoonTag nanobody from the surface of mammalian cells
198306pHCMV-SS-antiSunTag-TMDisplays anti-SunTag scFv from the surface of mammalian cells
198307pHCMV-SS-antiALFAtagNb-TMDisplays anti-ALFA nanobody from the surface of mammalian cells
198308pAWP78-PVCpnf_pvc12-HiBiTProduces PVCs tagged with HiBiT on the baseplate; used as assembly/loading control for payload loading assays
198309pAWP78-PVCpnfΔpvc15_pvc12-HiBiTNegative control for payload loading assay; produces PVCs deficient for payload loader (pvc15)
198310pBR322-Pdp1-HiBiTLoads HiBiT-tagged Pdp1 into PVCs; for payload loading assays; also contains Pnf
198311pBR322-Pdp1_aa1-289-HiBiTLoads a truncated payload; for identification of Pdp1 packaging domain
198312pBR322-Pdp1_aa1-190-HiBiTLoads a truncated payload; for identification of Pdp1 packaging domain
198313pBR322-Pdp1_aa1-133-HiBiTLoads a truncated payload; for identification of Pdp1 packaging domain
198314pBR322-Pdp1_aa1-76-HiBiTLoads a truncated payload; for identification of Pdp1 packaging domain
198315pBR322-Pdp1_aa1-38-HiBiTLoads a truncated payload; for identification of Pdp1 packaging domain
198316pBR322-Pdp1_aa39-331-HiBiTLoads a truncated payload; for identification of Pdp1 packaging domain
198317pBR322-Pdp1_NTD-GFP-HiBiTLoads HiBiT-tagged GFP into PVCs using Pdp1 packaging domain; also contains Pnf
198318pBR322-Pdp1_NTD-Cre-HiBiTLoads HiBiT-tagged Cre into PVCs using Pdp1 packaging domain; also contains Pnf
198319pBR322-Pdp1_NTD-ZFN_L-HiBiTLoads HiBiT-tagged L fragment of a zinc finger nuclease specific for human AAVS1 locus using Pdp1 packaging domain
198320TwinStrep-SUMO-Pdp1For affinity purification of Pdp1 payload
198321TwinStrep-SUMO-Pnf For affinity purification of Pnf payload
198448pAWP78-PVCpnf_Δpvc10Produces PVCs containing truncated spike tip protein (Pvc10).
198449pAWP78-PVCpnf_pvc13-Ad5RGDPK7_Δpvc10Produces PVCs retargeted with expanded tropism hAd5 knob (RGD and PK7 insertions); also contains truncated spike tip protein (Pvc10).
198450pAWP78-PVCpnf_pvc13-antiMouseMHCIINb_Δpvc10Produces PVCs retargeted with VHH7 anti-mouse MHC class II nanobody; also contains truncated spike tip protein (Pvc10).

Antibodies from Article